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Biomedical subjects

R A Good

Publications and source records attributed to R A Good.

At least 253 records · Page 14Linked to original sources
PubMed1989

Recombinant interleukin 2 therapy in severe combined immunodeficiency disease.

Severe combined immunodeficiency disease (SCID) is a congenital disorder of severe B- and T-lymphocyte dysfunction in which several pathogenic mechanisms have been identified. The present study describes a female child with SCID who had a primary defect in the ability of T cells to secrete interleukin 2 (IL-2). B- and T-cell numbers were normal, but their functions were severely deficient. Mitogen and antigen-driven lymphoproliferative responses were diminished but were correctable in vitro with recombinant IL-2 (rIL-2). The patient's phytohemagglutinin-stimulated lymphocytes expressed IL-2 receptors normally. Despite the presence of the gene for IL-2, the patient's cells were grossly deficient in messenger RNA for IL-2 and endogenous IL-2 production. Pokeweed mitogen-driven B-cell differentiation was decreased and was not corrected by the addition of normal T cells to the B cells. Two attempts at immune reconstitution by haploidentical bone marrow transplantation failed. Therapy with rIL-2 (30,000 units/kg, given daily i.v.) resulted in marked clinical improvement as well in improved T-cell functions. The child, now 3 yr old, has been on rIL-2 therapy for 2 yr and receives rIL-2 (30,000 units/kg) three times weekly at home. This case study points to a new direction in the treatment of such disorders with rIL-2.

B-Lymphocytes↗
PubMed1989

Positive selection of a T-cell subpopulation in the thymus in which it develops.

In SWR mice the expression with high-density V beta 17a (high V beta 17a) of the T-cell antigen receptors correlates with the CD4+8- subpopulation of thymocytes. By contrast, in thymocytes of SJL mice the expression of high V beta 17a is observed on the CD4+8- or CD4-8+ subpopulation. However, when the thymocytes from SWR mice have been developed in the SJL or B10 thymus but not in the H-2 compatible DBA/1 thymus, a greater proportion of thymocytes that express high V beta 17a was found to be CD4-8+. By contrast, only a small proportion of KJ23a+ thymocytes from SJL mice that had differentiated in the thymus of SWR or DBA/1 mice was CD4-8+, whereas a high proportion of CD4+8- cells expressed V beta 17a. Further, an intermediate proportion of KJ23a+ thymocytes that had derived from SJL donor mice was present on CD4-8+ thymocytes that had developed in B10.A(4R) thymus. These findings demonstrate that the appearance of a particular subpopulation of thymocytes (CD4-8+ with a beta chain of T-cell antigen receptor identified as V beta 17a) is determined by the histocompatibility complex products that are expressed in the thymic microenvironment in which the T cells develop.

Animals↗
PubMed1989

Decrease by chronic energy intake restriction of cellular proliferation in the intestinal epithelium and lymphoid organs in autoimmunity-prone mice.

In previous studies we demonstrated that chronic energy intake restriction (CEIR) by a diet relatively low in fat, relatively high in carbohydrate, and reduced 40% in total calories extends life span and delays development of autoimmune disease in autoimmunity-prone mice. To investigate a possible cellular basis for this dramatic action of CEIR, we analyzed the rate of incorporation of [3H]thymidine by cells of the intestinal epithelium, thymus, spleen, and mesenteric lymph nodes in ad libitum-fed mice vs. CEIR mice of three autoimmunity-prone strains. In New Zealand Black (NZB), MRL/MP-lpr/lpr (MRL/lpr), and BXSB mice, CEIR slowed the rate of uptake of [3H]thymidine and, by inference, the rate of cellular proliferation among epithelial cells along the entire length of the gastrointestinal tract. Furthermore, CEIR decreased the apparent proliferative rate of lymphoid cells of the thymus, spleen, and mesenteric lymph nodes. This action by CEIR on the proliferative rate of cells of these rapidly replicating cell populations may point to an important mechanism by which calorie restriction inhibits the development of autoimmune disease and extends longevity in autoimmunity-prone mice.

Animals↗
PubMed1989

In vitro synthesis of human immunodeficiency virus-specific antibodies in peripheral blood lymphocytes of infants.

An assay system was developed for the analysis of antibodies secreted in vitro against human immunodeficiency virus (HIV) by cultured peripheral blood lymphocytes of HIV-infected individuals. Cultures of peripheral blood lymphocytes were established with medium alone or with medium containing Epstein-Barr virus (EBV) or pokeweed mitogen. HIV antibodies were determined by an ELISA performed with commercial kits in which a whole virus extract served as antigen. Optimal antibody secretion was detected in 7-day peripheral blood lymphocyte cultures to which EBV had been added to provide polyclonal B-cell activation. Pokeweed mitogen-induced antibody secretion and spontaneous antibody secretion were less consistent. With EBV as a stimulus, the sensitivity and specificity of this assay for determining HIV infection status were each 100% in adults. When the assay was applied to infants and children, 23 of 24 symptomatic HIV-seropositive children (class P-2) and 11 of 33 asymptomatic seropositive infants aged less than or equal to 15 months (class P-0) tested positive for EBV-induced antibody secretion. Six of the 11 P-0 patients who tested positive have progressed to develop symptomatic disease, while the remainder are still seropositive at ages 2-15 months. Of the infants who were negative in this assay, all have remained asymptomatic. Treatment with 3'-azido-3'-deoxythymidine in infected adults and children has resulted in transient suppression of the in vitro antibody response in some instances. Thus EBV-induced synthesis of HIV-specific antibodies in vitro is a sensitive and specific indicator of HIV infection and is of help in determining infection status of asymptomatic seropositive infants who are classified as having "indeterminate" infection.

Acquired Immunodeficiency Syndrome↗
PubMed1989

Molecular heterogeneity of gamma delta T-cell antigen receptors expressed by CD4- CD8- T-cell clones from normal donors: both disulfide- and non-disulfide-linked receptors are delta TCS1+.

We investigated the molecular heterogeneity of gamma delta T-cell antigen receptors (TCR) expressed on T-cell clones generated from peripheral blood lymphocytes of normal donors. Extensive molecular heterogeneity was seen at the gamma-chain level and, to a lesser extent, at the delta-chain level. Both disulfide and non-disulfide gamma delta TCR were found and use different gamma chains with similar molecular masses (range, 41-43 kDa). In contrast, gamma chains of 55-60 kDa, which are expressed on T-cell lines derived from the peripheral blood of patients with immunodeficiency disorders, were not found on T-cell clones derived from the peripheral blood of normal donors. delta chains expressed on these T-cell clones had a molecular mass of 37 kDa and were either disulfide or nondisulfide linked. Significant delta-chain heterogeneity was identified in these clones using the anti-delta TCS1 and the anti-TCR delta 1 monoclonal antibodies. All clones tested were TCR delta 1+, whereas only 25% of the clones were delta TCS1+. The anti-delta TCS1 monoclonal antibody stained and immunoprecipitated both disulfide- and non-disulfide-linked gamma delta TCRs from different T-cell clones from normal donors.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1989

Influence of chronic energy intake restriction on intestinal alkaline phosphatase in C3H/Bi mice and autoimmune-prone MRL/lpr,lpr mice.

The influence of chronic energy intake restriction (CEIR) on the level and activity of intestinal alkaline phosphatase was investigated in mice of the autoimmune-prone MRL/lpr,lpr strain and in mice of the autoimmune-resistant C3H/Bi strain. In both strains of mice, CEIR of 40% resulted in a significant increase in intestinal alkaline phosphatase (IAP) specific activity in MRL/lpr,lpr mice after 10 wk of feeding, and in C3H/Bi mice after 6 wk of feeding. An increase in the amount of immunoreactive alkaline phosphatase antigen was also found to be associated with the increased enzyme activity in CEIR mice. These results suggest that a specific induction of an intestinal enzyme occurs or, alternatively, that there is a specific relative decrease in synthesis of intestinal proteins other than IAP as a function of CEIR. Thus, CEIR appears to regulate the expression of proteins in the small intestine in a specific manner.

Alkaline Phosphatase↗
PubMed1989

Acceptance of murine thyroid allografts by pretreatment of anti-Ia antibody or anti-dendritic cell antibody in vitro.

A new method for thyroid allografts in mice was established. Thyroids of C67BL/6J mice were treated with collagenase, and the follicles were isolated using a Percoll density gradient technique. These follicles were treated with anti-Ia antibody (Ab) or anti-DC Ab plus complement in order to eliminate DCs. The follicles were then mixed with agarose and transplanted under the left renal capsule of BALB/c mice. One hundred days after transplantation, acceptance of the grafts was verified by both histological study and the incorporation of 125I into the grafts. Allografts treated with C were rejected, whereas allografts treated with Ab plus C were accepted. When nontreated thyroids of C57BL/6J mice were grafted under the right renal capsule of BALB/c mice that had accepted DC-depleted thyroids of C57BL/6J mice, the nontreated thyroids were rejected. These findings indicate that DCs play a crucial role in the rejection of mouse thyroid allografts, and that the depletion of DCs permits allografts to be accepted without inducing donor-specific tolerance. Our method presented here may be developed as a viable strategy for the treatment of patients with congenital or acquired hypothyroidism.

Animals↗
PubMed1989

Strain differences in the early development of the thymus-dependent cells: precocity of T lineage cells in AKR mice as compared to those in C3H mice.

Early development of T lineage cells were compared between AKR and C3H mice by using two experimental strategies--neonatal thymectomy (NTx) and bone marrow transplantation (BMT)--between these two strains of mice. After NTx, AKR mice developed less wasting disease and showed better maintenance of several T cell functions. In addition, the response of neonatal spleen cells to PHA and ConA was much greater in AKR mice than in C3H mice. Further, when AKR mice were used as recipients of BMT, cell numbers recovered from thymuses between 2 and 7 weeks after reconstitution were consistently much greater (about 10 times greater) than those from chimeras where C3H mice were used as recipients, regardless of the donor strains of bone marrow cells. However, 4 weeks after BMT the proliferative responses to ConA were consistently higher in the donor-derived thymocytes from chimeras where AKR mice were used as bone marrow donors than in those from chimeras in which C3H were donors. The present findings suggest that these differences may be attributed to characteristics of recipient microenvironment (e.g., thymic stroma) which maintain developing thymocytes and supply them to the peripheral lymphoid tissue. Alternatively the differences may to some degree also be attributable to characteristics of the thymic progenitors themselves, which may determine the rates of maturation of thymocyte functions.

Animals↗
PubMed1989

A study on graft-versus-host reaction (GVHR) by Simonsen's splenomegaly assay. Cells and antigen systems involved in induction of GVHR.

Cells and histocompatibility antigen systems involved in graft-versus-host reactions (GVHR) were analyzed using Simonsen's splenomegaly assay employing various combinations of donor and F1 hybrid recipients mice. Most of the cells proliferating in spleens of mice undergoing GVHR were J11d+, and had histological features of cells of the hematopoietic lineage. The proportions of CD3+ T cells were decreased in the spleens. Disparity at minor histocompatibility determinants of AKR, I-E and H-2D regions between B10.A(4R) donors and (4R X AKR) F1 recipients evoked only negligible GVHR. On the contrary, disparity at H-2K and/or I-A regions appeared to be sufficient to permit induction of full GVHR. When surface markers of donor spleen cells were analyzed, it was shown that Thy-1+ and/or MEL-14+ cells caused a strong effect on GVHR. Further, either CD4+ or CD8+ T cell subset could induce significant GVHR. However, synergistic influences of these two T cell subsets on one another in GVHR were observed. The present results raise the possibility of using Simonsen's assay along with a number of reagents to identify the contribution of subsets of T lymphocytes and in analyzing precise contributions of cellular components from both donor and recipient, and also of the target antigen systems of the recipient that contribute to early events involved in GVHR.

Animals↗
PubMed1989

Disulfide-linked gamma delta T cell antigen receptors expressed on T cells derived from patients with primary immunodeficiency disorders.

T cell lines or clones from two patients, one with a partial DiGeorge syndrome and one with severe common variable immunodeficiency expressed disulfide-linked gamma delta T cell antigen receptor (TCR) comprised of a gamma-chain polypeptide of 40-43 kD, and a delta-chain polypeptide of 37-40 kD. This gamma delta TCR appears to be similar to that found on T cell clones, and lines derived from peripheral blood lymphocytes from normal donors. Previous studies have shown that T cell lines derived from the peripheral blood of patients with immunodeficiency disorders express non-disulfide-linked gamma delta TCR. In contrast to the latter and coincident with findings in the present study, the vast majority of T cell lines and clones derived from the peripheral blood of normal donors express disulfide-linked gamma delta TCR.

Antigens, CD↗
PubMed1989

Graft failure following bone marrow transplantation for severe aplastic anemia: risk factors and treatment results.

Graft failure was analyzed in 625 patients receiving allogeneic bone marrow transplants from HLA-identical sibling donors as treatment for severe aplastic anemia. Sixty-eight (11%) had no or only transient engraftment. Second bone marrow transplants were successful in achieving extended survival in 16 of 27 patients with transient initial engraftment but in none of ten patients with no sign of engraftment after the first transplant. The major factors associated with a reduced risk of graft failure were use of radiation for pretransplant immunosuppression and use of cyclosporine rather than methotrexate or T-cell depletion of the donor bone marrow for prophylaxis against graft-v-host disease (GVHD). Among 266 patients prepared for transplantation with cyclophosphamide alone, the risk of graft failure was increased in patients who received previous transfusions and reduced in those who received corticosteroids for previous therapy. Neither cell dose nor administration of donor buffy coat cells affected the probability of engraftment. Although use of radiation in conditioning reduced graft failure, survival was not improved. Posttransplant treatment with cyclosporine and avoidance of pretransplant blood transfusions were associated with improved survival.

Actuarial Analysis↗
PubMed1989

A study on type II collagen-induced arthritis in allogeneic bone marrow chimaeras.

In the present investigation, the incidence, day of onset and severity of type II collagen (IIC)-induced polyarthritis (CA) in allogeneic bone marrow chimaeras were compared. These chimaeras were prepared by using reciprocal combinations between DBA/1J (H-2q, Igh-1c, Hc1), which are highly susceptible to CA, and SWR/J (H-2q, Igh-1c, Hc0), which are CA resistant, strains of mice. The four kinds of chimaeras, [SWR----DBA/1], [DBA/1----DBA/1], [DBA/1----SWR] and [SWR----SWR], were shown to be full chimaeras. [SWR----DBA/1] and [DBA/1----DBA/1] chimaeras showed serum C5 antigens and most of these mice developed CA, even though severity of CA in [SWR----DBA/1] chimaeras was less than that in [DBA/1----DBA/1] syngeneic chimaeras. By contrast, [DBA/1----SWR] and [SWR----SWR] chimaeras completely lacked serum C5 and did not develop CA at all. However, most of these chimaeras produced considerable total IgG and IgG2a antibodies to IIC in the sera after immunization with the antigens, although differences in the serum levels of IgG2a antibodies were seen between [SWR----DBA/1] and [DBA/1----SWR] allogeneic chimaeras. The present results demonstrate directly a crucial role of complement cascade in the induction and development of CA and reveal that lymphoid cells of SWR mice potentially have the ability to develop CA. Differences in severity of the CA observed in some of these chimaeras, however, could not be readily explained by differences in C5 concentrations alone.

Animals↗
PubMed1988

Bone marrow cells from allogeneic bone marrow chimeras inhibit the generation of cytotoxic lymphocyte responses against both donor and recipient cells.

When added to a mixed lymphocyte culture, bone marrow cells suppress the generation of CTL activity against H-2 Ag shared by the BM cells and the stimulator cells. These cells have been referred to as veto cells and are thought to play a role in maintaining self-tolerance. We analyzed the H-2 specificity of the suppression expressed by the veto cells from H-2 incompatible bone marrow chimeras, because lymphocytes of such chimeras had been shown to be tolerant to both donor and recipient Ag when tested by CTL responses. We found that the bone marrow cells of such chimeras which were featured by non-T and non-B cell characteristics inhibited the generation of CTL directed against either donor or recipient Ag, but not against third-party Ag. These observations suggest that in allogeneic chimeras the veto or veto-like cells alter the inhibitory specificity exhibited in the recipient microenvironment and indicate that these cells are directly involved in the induction and maintenance of self-tolerance.

Animals↗
PubMed1988

Human IFN-gamma production is inhibited by a synthetic peptide homologous to retroviral envelope protein.

A synthetic 17 amino acid peptide (CKS-17) homologous to a highly conserved region of human and animal retroviral transmembrane proteins was investigated for its influence on the in vitro production of IFN-gamma from human peripheral mononuclear cells. The results showed that CKS-17 coupled to a carrier protein, BSA, inhibited production of IFN-gamma in a dose-dependent manner. Controls, consisting of BSA, which had undergone the coupling procedure or neurotensin coupled to BSA in an identical manner as CKS-17, showed no such inhibition. Reduction in IFN-gamma production could not be attributed to decreased viability of cells, delay of IFN-gamma production or to involvement of suppressor cells. Moreover, inhibition of IFN-gamma production was not related to the inhibition of DNA synthesis. The inhibition appeared to be a direct effect of CKS-17 on IFN-gamma-producing cells. Kinetic studies revealed that this suppression occurred when CKS-17 was introduced to the culture concurrent with or within 48 h after introduction of IFN inducers. Preincubation experiments showed that the presence of CKS-17 in the culture medium was not necessary to exert its inhibitory effect. These results suggest that a portion of retroviral envelope proteins possess important immunomodulatory actions.

Amino Acid Sequence↗
PubMed1988

The presence of autoantibodies specific for NZB serum factors in adult NZB mice and the establishment of monoclonal autoantibodies against these humoral factors.

Humoral factors in serum of young NZB mice enhance maturation of B-lymphocyte precursors in vitro. A blot ELISA assay identified autoantibodies against the serum factors. NZB-SFs (designated NZB-SF alpha, pI 3.5-4.0, and NZB-SF beta, pI 7.8) were purified by sequential steps. Both had a molecular weight (MW) of approximately 23,000 in SDS-PAGE. NZB mice develop autoantibodies against NZB-SFs by 2 months of age; titers increased progressively with age. Non-autoimmune-prone mice did not produce autoantibodies against NZB-SFs. We then developed two hybridoma clones, IIC1C1 and IIC1M4, which produce monoclonal autoantibodies against NZB-SF alpha and NZB-SF beta, respectively. Both IgM autoantibodies could be affinity purified with a column of CNBr-Sepharose 4B gel conjugated with anti-mouse IgM antibody. Neither IIC1C1 nor IIC1M4 abolished bioactivity of recombinant mouse IL-1 alpha, human IL-1, mouse, rat, or human IL-2, mouse IL-3, or colony-stimulating factor. Neither antibody reacted to recombinant mouse IL-1 alpha, IL-4, TNF alpha, or IFN gamma in blot ELISA assays. Monoclonal autoantibodies IIC1C1 and IIC1M4 were used to purify NZB-SFs. SDS-PAGE of the affinity-purified NZB-SFs revealed bands of 23 and 60 kDa, and proteins extracted from the bands were reactive to our monoclonal autoantibodies.

Animals↗
PubMed1988

A surgical technique for experimental free skin grafting in mice.

In 3031 inbred mice, 3093 allogeneic free skin grafting procedures, including 62 double grafts, were performed using trunk skin as the graft tissue. We developed a new method of graft bed preparation, using corneal scissors for the incision of the square frame of the graft bed and gauze sponges held in a pair of mosquito forceps for the blunt dissection of recipient skin. This method effectively preserved the panniculus carnosus of the graft beds. Grafts from the donor trunk skin were also prepared by a new method in which the skin was spread on aluminium foil, cleaned of superfluous tissues using large gauze sponges held in forceps, cut into appropriately sized grafts with a scalpel, and marked by scoring in order to designate the original direction of the grafts. The graft was approximated to the graft bed by 8 interrupted sutures of 5-0 silk, then covered with a bandage and protective tape for 7 days. The overall operative and postoperative mortality, in most cases attributable to overdoses of anesthetic drugs, was 3.8 per cent (114/3031) and the overall graft failure rate, in most instances due to incorrect covering of the grafts, infection or inside-out-grafting, was 1.2 per cent (36/2979).

Animals↗
PubMed1988

Guinea pig and gerbil erythrocytes rosette with different cells in the blood, bone marrow, and thymus of the cat.

Cat thymocytes, bone marrow cells, and peripheral blood leukocytes (PBL) formed rosettes with guinea pig (GP) and gerbil (G) erythrocytes (E). In PBL from adult cats the frequency of rosettes was 27% with GPE and GE, while an average of 33% bone marrow cells formed rosettes with GPE and only 4% with GE. Thymocytes from kittens showed a high percentage of rosettes with both GPE and GE (35 to 81%), with the frequency of each type varying with the thymus tested. Fluorescein isothiocyanate labeling of one of the erythrocyte species revealed these cells to be rosetting with different nucleated cells; i.e., a low percentage (3-5%) of the rosettes formed with PBL and bone marrow had both labeled and unlabeled erythrocytes. In contrast, "mixed" rosettes were observed with a significant number of thymocytes, averaging 33% of thymocytes from six animals. A further distinction between the GE- and GPE-rosetting cells was revealed by a monoclonal antibody which blocked GE rosette formation without interfering with the binding of GPE to PBL and thymocytes. PBL could be depleted of either GPE- or GE-rosetting cells, with retention of IgG+ cells and cells capable of rosetting with the second erythrocyte species in the nonrosetting fractions. Stimulation of the latter nonrosetting fractions with pokeweed mitogen for induction of Ig synthesis revealed a T-lymphocyte specificity of the GE- and GPE-rosetting cells. PBL depleted of GE-rosetting cells yielded an increased Ig production, two- to threefold above the control; in contrast, depletion of GPE-rosetting cells from PBL resulted in a failure of the remaining cells to respond. These results suggest that T-suppressor cells of the cat are contained in the GE-rosetting fraction and T-helper cells are rosetted with GPE.

Animals↗